Abstract Collaborative research was conducted at the INRA Research Centers to assess the microbial control potential of Beauveria bassiana- and Lecanicillium lecanii-based formulations against whiteflies in protected crops under Mediterranean conditions. Four series of small-scale glasshouse trials were performed in 1999 and 2000 in southern France. Two applications at 4–5 day intervals of Naturalis-L and Mycotal were conducted on young larvae of the greenhouse whitefly, Trialeurodes vaporariorum, at rates recommended by the manufacturers. Because of the expectation that environmental conditions prevailing in Mediterranean greenhouse crops may lead to greater climatic constraints for mycoinsecticide efficacy than in more temperate areas, manipulation of the greenhouse climate has been used to aim at optimizing mycoinsecticide efficacy. The climatic management strategy was mainly based on closing the ridge vents 2 h more at night-time in so-called “humid” glasshouse compartment than in a “dry” one. Thus, the daily period at high humidity (>90% RH) was two or three times longer in the “humid” compartment than in the “dry” one. In spite of this differential, mycoinsecticide treatments reduced numbers of surviving whitefly larvae by >85% in the “humid” compartment as expected as favorable, as well as in the “dry” compartment, expected as unfavorable. The results indicated clearly that both B. bassiana- and L. lecanii-based mycoinsecticides have a strong potential for microbial control of whitefly larvae infesting tomato crops at moderate ambient humidity in Mediterranean glasshouses. Our investigations provided strong arguments for explaining these unexpected results. The RH conditions prevailing in the targeted insect habitat should be greatly disconnected from that of the ambient glasshouse air. We suggest that strategies of mycoinsecticide optimization against phyllophagous insects in protected crops have to take into account factors acting on the leaf transpiration activity.